%0 Generic
%A Hernandez Bücher, Jochen Estebano
%C Heidelberg
%D 2025
%F heidok:35810
%R 10.11588/heidok.00035810
%T Design and development of synthetic immune cells for  biomedical applications
%U https://archiv.ub.uni-heidelberg.de/volltextserver/35810/
%X Cytotoxic lymphocytes are able to recognize malignant cells through surface specific receptors and eventually induce apoptosis in their targets. This is a crucial property of the mammalian immune system’s own vigilance capabilities, to ensure the integrity of the organism. However, cancer cells that are able to evade these complex immune regulatory mechanisms develop the ability to escape recognition and elimination. Therefore, adoptive cell therapeutic approaches, with the goal to restore the ability of cytotoxic lymphocytes to recognize these malignant cells again, have evolved as promising tools for cancer treatment.  Nonetheless, this cell-therapy implements top-down genetic engineering that has some limitations in terms of turn-over time, cost-effectiveness, versatility, and side effects. To reduce the complexity of living cells, I designed and developed in this thesis two bio-engineering approaches for the assembly of bio-inspired synthetic cytotoxic immune cells.   	First, I describe the bottom-up assembly of giant unilammelar vesicle (GUV) based synthetic immune cells which are able to induce apoptosis in target cells and at the same time to discriminate, on an antigen mediated basis, between two cancerous cell lines. Based on the achievements with the synthetic immune cells, I developed a second strategy to transfer the conceptual idea of the target cell-specific cytotoxicity to natural, red blood cell derived, membrane scaffolds. For this, I combined top-down and bottom-up synthetic biology approaches to develop cytotoxic cells on the basis of ghost red blood cells (GRBCs). Towards this end, I integrated functional lipids into the outer membrane of GRBCs and functionalized them with the apoptotic ligands FasL and TRAIL. Moreover, to maintain stable long-lasting linkage between ligands and GRBCs as required for in vivo applications, I implemented a bio-orthogonal functionalization strategy based on coordinative Co3+ complexation. Further, I was able to show that GRBC-based cell surrogates are not endocytosed and eliminated by macrophages. Importantly, in vivo experiments with mice revealed that the engineered GRBCs can circulate in the bloodstream for at least 24 h. This underscores the value of the developed system for future studies with synthetic cells in a pre- clinical context.  Therefore, this study paves the way for alternative cell-based immunotherapies for cancer treatment with potentially higher versality, lower cost and reduced side effects, providing an on-demand approach in personalized medicine.